Translating research into practice
Immunisation for genital viral infections
Ian Frazer, Research Director, The Translational Research Institute
Brisbane, Australia
Infection causes >20 % of cancer
And we’re still finding new ones.
%
Infectious agent
Associated Cancer
Viruses
5.2 Papillomavirus
Anogenital, oropharyngeal
3.9 Hepatitis B virus
1.0 Epstein Barr Virus
Hepatocellular
Nasopharyngeal Ca, Lymphoma
2.9
0.9
Hepatocellular Ca
Potentiates viral cancers
Hepatitis C virus
HIV
<0.2 HTLV -1
T cell leukemia
<0.2 Merkel polyomavirus Merkel’s disease
Other
<0.2 HHV -8
Kaposis sarcoma
5.5
Gastric
H. Pylorii
<0.2 Parasites
Bladder, ?HCC
NB Infections common, cancer rare consequence
Parkin DM et al Int J
Cancer 118, 3030 (2006)
Treating infections like HPV is a challenge
Preventing them seems more sensible
HPV vaccine development:
A 15 year process
Zur
Hausen
Cancer associated
Human Papillomaviruses
(~1980)
100% of cervical cancer
and 30% of head and
neck cancer caused by
“high risk” HPVs (16,18)
Jian Zhou
Virus Like particles1
(~1990)
Major L1 capsid
protein expressed in
vitro self assembles
to VLPs
1:J. Zhou, X. Y. Sun, D. J. Stenzel, and I. H. Frazer. Virology 185 (1):251-257, 1991.
HPV vaccines
(~2005)
- VLPs+ adjuvant
- Neutralising Ab
- Protection >8yrs
-Cervarix
- HPV 16, 18
-Gardasil
– HPV 6,11,16,18
Nine steps to translate HPV
research into practical outcomes
•
•
•
•
•
•
•
•
Define the problem – HPV and cancer
Develop the science –recombinant VLPS
Find commercial partners
Scale up the manufacture
Complete safety and efficacy trials
Educate the profession, govt, the public
Confirm field efficacy
Improve the product
– 9 valent vaccines, 2 dose regimens
• Encourage deployment globally
• Develop effective immunotherapy
Zur Hausen
Jian Zhou
Merck/GSK
Merck/GSK
Many groups
Ongoing
Disclosure of conflict of interest
Dr Ian Frazer and the University of Queensland
benefit financially from commercial sale of the
prophylactic HPV vaccines discussed in this talk
CONFIRMING FIELD
EFFICACY
Genital warts in young vaccinated women in
Australia
Ali, H. Et al BMJ 2013;346:f2032 doi: 10.1136/bmj.f2032
Geniital Warts have gone away in their unvaccinated
male partners
Ali, H. Et al BMJ 2013;346:f2032 doi: 10.1136/bmj.f2032
Differences in HPV genoprevalence between
prevaccine and postvaccine populations.
Latest Australian data for PAP samples
90% reduction in HPV16, 18 in < 30yrs
Tabrizi S N et al. J Infect Dis. 2012;infdis.jis590
ENABLING GLOBAL USAGE
There is disparity between HPV vaccine
programs and cervical cancer prevalence
http://www.indexmundi.com/blog/wp-content/uploads/2013/05/HPV-vaccine-infographic.jpg
Vaccine delivery logistics in Vanuatu:
staffing, and a cold chain
Vanuatu
->50 islands
~250,000 people
~25 doctors
-91 parliamentarians!
-1 vaccine fridge
-0 reliable electricity
500 well women over 30
Bx result
Number
Cancer
5
CIN 3
17
CIN 1
23
Education in Vanuatu: parents,
children, staff, and government
Parents and Kids under the Banyan Tree- north Efate
When vaccine is available,
vaccination programs in
subsistence economies can be
as successful as Australia
State
Age
Dose 1
Coverage (%)
Dose 1 immunised receiving
Dose 2 (%)
Dose 3 (%)
Australia 12-18 (Opt out)
~ 90%
96
86
Australia 18-25 (Opt in)
~80%
82
50
Vanuatu
~80%
98
93
10-12 (Opt out)
~80% coverage for bivalent HPV vaccine for 10-12 year olds in 2008, and 2013
Vaccine through Australian Cervical Cancer Foundation: delivery by government
15
Fighting genital cancer with immunity
The way of the future?
Cervical cancer prophylactic vaccines
are not therapeutic
Source : ACIP website 2008/03/30
Immunotherapy for HPV
associated cancer
• E6/E7
– recombinant bacterial
fusion of HPV16 E6 and
E7 proteins in 8M Urea
100nm
• ISCOMATRIX®
adjuvant 120µg/dose
– Quillaia saponin based
adjuvant
– typically 40nm cage like
structures
– promotes both humoral
and cell mediated
immunity
ISCOMATRIX® adjuvant
HPV 16 vaccine
Immunotherapy: can we get rid of
existing HPV infection
Immunology – a nice response
Antibody +++
DTH
+++
Cytotoxic +++
T cells
DTH (mm)
•
•
•
20mg (3x)
>200
100
50
40
40
30
30
20
20
10
10
0
0
200mg (1x)
>200
100
1 quadrant
9
2 quadrants 11
3+ quadrants 10
Frazer, I. H., M. Quinn, et al. (2004).
Vaccine 23(2): 172-181.
Placebo
(1x or 3x)
>200
100
50
50
40
40
30
30
20
20
10
10
0
Colposcopy- no change
Pre- and post- vaccination
60mg (3x)
>200
100
50
0
Histology- no change
Pre vaccination
histology
N
Post vaccination
histology
CIN
2,3
CIN 2,3
23
22
CIN 1
7
1
CIN
1
Nil
1
3
3
Modelling epithelial cancer immunotherapy
Keratinocyte-immune cell interactions
DONORS
K14 transgenic
RECIPIENT
Nontransgenic
control
Allo matched
nontransgenic
• Antigen expressed only in basal keratinocytes
• Stable expression of antigen over time without tumor
related phenomena
• Grafts expressing some antigens (eg OVA) reject
spontaneously,
• Grafts expressing others (eg HPV E6, E7) don’t
How does inflammation work?
A skin resident immunocyte locally inhibits E7
specific effector T cell function
immune cells
in E7 graft
No immune
cells (rag-/-)
in E7 graft
Mattarollo, S. R., A. Rahimpour, et al.
(2010). The Journal of Immunology 184(3):
1242-1250.
Not reproduced by:
Depletion of CD25+ regulatory cells from graft (Mab),
Elimination of FoxP3 positive cells from graft ( DEREG mouse)
Therefore – another regulatory T cell population exists in skin
Steve Mattarollo, Rachel DeKluyver, Christina Gosmann
NKT CELLS AND IFN-γ ARE
KEY REGULATORS OF LOCAL
T CELL FUNCTION
The inhibitory cell is an NKT cell.
NKT cells
No NKT cells
Note – inhibition of
rejection local to NKT
replete graft
Conclusion: NKT cells locally inhibit effector function
of cytotoxic T cells
Mattarollo, S. R., A. Rahimpour, et al. (2010). "Invariant NKT cells in hyperplastic skin induce a local immune
suppressive environment by IFN-gamma production." The Journal of Immunology 184(3): 1242-1250.
Footnote:
Of mice and (wo)men!
Parameter
In mice
In humans
IDO
 In E7tg + Inhibits rejection
 In CIN
a
IL-17
 In E7tg + Inhibits rejection
 In CIN
a
IFN-g
 In E7tg + Inhibits rejection
 In CIN
a
Mast cells
 In E7tg + Inhibits rejection
 In CIN a
IL1Ra
 In E7tg + Inhibits rejection
 In CIN b
NKT cells
Inhibit rejection
a: Frazer Lab Data
b: Fujiwaki R Gynecol Oncol. 2003 Apr;89(1):77-83
Application of a new vaccine technology
to an old problem
Progress on a herpes vaccine
Conventional vaccines
haven’t worked?
N Engl J Med. 2012 Jan 5; 366(1): 34–43
Modified codon usage allows viral
gene expression in cell lines
Wt BPVL1
HB BPVL1
Wt BPVL2
HB BPVL2
Using polynucleotides as vaccines
Making the body’s cells do the work
Codon modification = better responses
IMMUNE RESPONSE
TO VACCINE
1.5
CODON MODIFIED (1 shot)
CODON MODIFIED (2 shots)
1.0
NO MODIFICATION
0.5
0.0
1:50
1:200
1:800
Serum Dilution
1:3200
Another improvement
Ubiqitination = better cellular immunity
H6L1 E7
UH6L1E7
L 1 mod
Ubi
E7
L 1 mod
E7
75
50
25
0
A
MIX (LN)
MIX (S)
Control (LN)
Control (S)
1.2
B
1.0
0.8
0.6
0.4
0.2
0.0
1:50
Ab to
denatured L1
IFN spots/ 106 cells
CTL
100
Conformational
antibodies
Mixed polynucleotide vaccines have
the immunogenicity of both
components
1:200
1:800
1:3200
1.2
C
1.0
0.8
0.6
0.4
0.2
0.0
1:50
1:200
1:800
1:3200
HSV- 2 Vaccine Development
o Based on glycoprotein D
o Initial gD2 constructs:
• control (wt CU gD2)
• codon modified (O1, O2 and O3 gD2)
• codon de-optimised (W gD2)
o Initial studies identified O2 as best performer
o Made ubiquitinated version of O2
Challenge Survival
survival (%)
50 X LD 50
100
90
80
70
60
50
40
30
20
10
0
500 X LD 50
100
90
80
70
60
50
40
30
20
10
0
TKO2-gD2
gD mix
O2-UgD2 25-331
W-gD2
empty vector
0 1 2 3 4 5 6 7 8 9 101112131415161718192021
0 1 2 3 4 5 6 7 8 9 101112131415161718192021
days post-infection
days post-infection
HSV-2 Vaccine Phase 1 Study
o Safety, Tolerability & Immunogenicity Endpoints
o Open Label, Multiple Dose Escalation
o Doses of 10µg, 30µg, 100µg, 300µg, and 1mg
o Intradermal injections (Days 0, 21 and 42)
o 22 healthy sero-negative 18-45 yr old subjects
enrolled from 59 volunteers
HSV immunogenicity studies
4
10
30
100
300
1000
3
2
1
DTH
DOSE: mg
DTH REACTION SCORE
DOSE: mg
15
10
30
100
300
1000
10
5
week7
week10
TIME POINT
Week
24 h
48 h
ee
k1
0
week4
w
0
w
ee
k7
0
w
ee
k4
New reactive gD peptides
IFN-g Elispot data
HSV-2 Vaccine Phase 1b Study
o 40 18-45 yr old subjects with recurrent culture/PCR
proven HSV-2
oRandomised, blinded, 1mg Dose or placebo
oIntradermal injections (Days 0, 21 and 42)
oSafety, Tolerability & Immunogenicity primary
endpoints
oViral shedding before (2 months) and after (2 months)
vaccine secondary endpoint
Acknowledgements
• Frazer Lab, Diamantina Institute, TRI, Brisbane
• (Especially Christina Gosmann, Dr Steve Mattarollo, Dr
Deepak Mittal, Dr Antje Blumenthal )
• Lambert Lab, McArdle Ins
FUNDING
• NHMRC Australia
• NCI, NIH
• Cancer Council Australia
• Australian Cancer Research Foundation